NOD2 CRISPR Activation Plasmid (h): sc-400668-ACT

Human NOD2 (nucleotide-binding oligomerization domain-containing protein 2) is a cytosolic pattern-recognition receptor that detects bacterial peptidoglycan fragments such as muramyl dipeptide and initiates innate immune signaling. Upon activation, NOD2 engages RIPK2 to stimulate NF-κB and MAPK pathways, shaping inflammatory gene expression, antimicrobial responses, and epithelial barrier homeostasis. NOD2 also interfaces with autophagy-related processes and microbial sensing in myeloid and intestinal epithelial cells. Genetic and functional perturbations in NOD2 signaling are linked to dysregulated host–microbe interactions and inflammatory phenotypes, supporting its study in mucosal immunity and infection-associated inflammation models.

NOD2 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous NOD2 expression without altering the underlying DNA sequence.

NOD2 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the NOD2 locus in human cell lines. The system is built around a catalytically inactive Cas9 (dCas9) carrying two inactivating mutations (D10A and N863A) that eliminate nuclease activity while preserving DNA binding. This dCas9 is fused to VP64, a potent transcriptional activator, and is co-expressed with a blasticidin resistance gene for selection. The second plasmid encodes the MS2-p65-HSF1 fusion protein, a secondary activator complex that works in concert with dCas9-VP64, alongside a hygromycin resistance gene. The third plasmid encodes a target-specific 20 nt sgRNA fused to two MS2 RNA aptamers that recruit the MS2-p65-HSF1 complex to the activation site, accompanied by a puromycin resistance gene. The three plasmids are delivered at a 1:1:1 mass ratio for balanced expression of all system components.

Once assembled at the target locus, the SAM complex binds within approximately 200 bp upstream of the NOD2 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous NOD2 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native NOD2 locus and enabling the study of NOD2-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of NOD2 pathway restoration in tumor cells with silenced or reduced NOD2 expression.

For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.